Hassouneh Wafa, Christensen Trine, Chilkoti Ashutosh
Duke University, Durham, North Carolina.
Curr Protoc Protein Sci. 2010 Aug;Chapter 6:6.11.1-6.11.16. doi: 10.1002/0471140864.ps0611s61.
This unit presents a recombinant protein purification method that employs an elastin-like polypeptide (ELP) as a purification tag. ELPs undergo a sharp and reversible phase transition when heated above their lower critical solution temperature. ELPs retain this behavior when they are fused to a protein, and thereby provide a simple method to isolate a recombinant ELP fusion protein from cell contaminants by cycling the solution through the insoluble and soluble phase of the ELP fusion protein using a procedure that is termed Inverse Transition Cycling. This method does not require the use of chromatography, so it is cost-effective, easy to scale up, and easy to multiplex.
本单元介绍了一种重组蛋白纯化方法，该方法采用弹性蛋白样多肽（ELP）作为纯化标签。当加热到其低临界溶液温度以上时，ELP会发生急剧且可逆的相变。当ELP与蛋白质融合时，它们会保留这种特性，从而提供一种简单的方法，通过使用称为反向转变循环的程序，使溶液在ELP融合蛋白的不溶相和可溶相之间循环，从细胞污染物中分离重组ELP融合蛋白。该方法不需要使用色谱法，因此具有成本效益，易于扩大规模且易于多路复用。
